| 摘要 |
PURPOSE:To obtain the subject new polypeptide having a specific amino acid sequence, useful for detecting antigens for new non-A-and-non-B hepatitis virus having been unable to be detected by conventional methods, thus usable to diagnose non-A-and-non-B hepatitis. CONSTITUTION:The serum of patients with non-A-non-B hepatitis and negative in HBs antigen and HBV-DNA is pooled; polyethylene glycol is added to the pool to effect precipitation, and viral RNA is then extracted from the resultant precipitate by e.g. guanidine-thiocyanate method, a c-DNA is synthesized using this viral RNA as template and then incorporated in a phage vector to constitute a cDNA library. This library as probe is then screened using a synthetic polynucleotide composed of the 8 bases derived from the complement chain of non-A-and-non-B hepatitis virus polynucleotide followed by restriction enzyme treatment of the DNA of positive clone to bind this DNA to a manifestation vector, and the DNA is manifested in host cells, thus obtaining the objective non-A-non-B hepatitis virus antigen polypeptide of the formula (X is such that the N-terminal is H, Met-Ala or Ala). |
